Particle arrangement method for multiple biochemical detection

A biochemical detection and microparticle technology, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of accurate calculation of light intensity, reduced sensitivity of detection methods, low detection efficiency and false positives, etc., to achieve convenient operation and improve detection Efficiency and detection throughput, the effect of solving optical detection interference

Pending Publication Date: 2020-12-22
WUHAN TEXTILE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

The disadvantage of this chip is that the microspheres are fixed on the step boundary, which easily leads to the blockage of the microchannel; especially in multiple biochemical detection, the more types of microspheres are used, the greater the possibility of the fluid channel being blocked, which may cause Affect hybridization efficiency, resulting in reduced sensitivity of detection method
Furthermore, in optical detection, the structural change of the step chip at the step will lead to optical interference such as refraction and shadows in the process of optical detection, which will cause deviations in the accurate calculation of light intensity in the later detection process
Finally, the detection of the target object by microspheres in the microfluidic chip needs to ensure sufficient contact between the two. Therefore, the multiple biochemical detection method of immobilizing the microspheres first and then performing hybridization may easily lead to problems such as low detection efficiency and false positives.

Method used

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  • Particle arrangement method for multiple biochemical detection
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Examples

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Effect test

Embodiment 1

[0017] A Microparticle Array Method for Multiplexed Biochemical Detection of Three Cytokines. In this example, the microfluidic chip is prepared with a glass cover slip and a substrate, the angle between the glass cover slip and the substrate is 0.01 degrees, and among the N kinds of immunoparticle complexes used in conjunction, N=3 , the particles become spherical, specifically: immune particle complexes with a size of 1 μm that capture the cytokine IL-10, immune particle complexes with a size of 1.5 μm that capture the cytokine IFN-γ, and immune particle complexes with a size of 2 μm that capture cells Immune microparticle complexes of factor TNF-α. Specifically include the following steps:

[0018] (1) Preparation of a microfluidic chip with an angle of 0.01 degrees: punch holes at 16 mm and 5.5 mm from the edges of the cover and substrate respectively, then clean the cover and stick a layer of mask on its upper surface, A portion of about 16 mm from one end of the cover ...

Embodiment 2

[0026] An alignment method for eight large-size particles. In this embodiment, the microfluidic chip is prepared by using a glass cover slip and a substrate, and the angle between the glass cover slip and the substrate is 0.1°. The particles of N kinds of sizes used together, N=8, the particles are spherical, specifically: microspheres with a size of 15 μm, microspheres with a size of 20 μm, microspheres with a size of 25 μm, and microspheres with a size of 30 μm. 35 μm microspheres, 40 μm size microspheres, 45 μm size microspheres, and 50 μm size microspheres. Specifically include the following steps:

[0027] (1) Preparation of a microfluidic chip with an angle of 0.1°: punch holes at 40 mm and 0.6 mm from the edges of the cover and substrate, respectively, and then clean the cover and stick a layer of mask on its upper surface, A part of about 40 mm from one end of the cover slip was exposed by laser etching, and then a layer of PDMS prepolymer with a thickness of 60 μm w...

Embodiment 3

[0034] A Microparticle Array Method for Multiplexed Biochemical Detection of Three Viruses. In this embodiment, the microfluidic chip is prepared with a glass cover slip and a substrate, the angle between the glass cover slip and the substrate is 0.02 degrees, and among the N types of immunoparticle complexes used in conjunction, N=3 , the particles are spherical, specifically: immune particle complexes with a size of 7 μm that capture influenza A virus, immune particle complexes with a size of 9.5 μm that capture influenza B virus, and immune particle complexes with a size of 10.5 μm that capture syncytia Virus immune particle complexes. Specifically include the following steps:

[0035] (1) Preparation of a microfluidic chip with an angle of 0.02 degrees: punch holes at 15 mm and 8.6 mm from the edge of the cover and substrate respectively, then clean the cover and stick a layer of mask on its upper surface, A part of about 15 mm from one end of the cover slip was exposed ...

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Abstract

The invention discloses a particle arrangement method for multiple biochemical detection. According to the method, a micro-fluidic chip and at least N immune particle complexes with different sizes are specifically utilized, and N is an integer greater than 2; and solutions mixed with N immune particle complexes with different sizes are injected into a wedge-shaped inner cavity of a micro-fluidicchip from a fluid inlet of the micro-fluidic chip at a flow speed of 50-300 [mu]L/min, and the N immune particle complexes with different sizes are naturally arranged in the inner cavity of the micro-fluidic chip respectively under the action of a fluid. The method is simple and easy to operate, a detection result is accurate, sensitive and free of optical interference, the chip is not prone to blockage, multiple encoding can be achieved, and the method is widely applied to detection of biochemical markers such as protein, nucleic acid and pathogenic bacteria in the fields of clinical diagnosis, biochemical analysis and drug screening and has wide application prospects.

Description

technical field [0001] The invention relates to a particle array method for multiple biochemical detection. The invention can be widely used in the detection of biochemical markers such as proteins, nucleic acids, and pathogenic bacteria in the fields of clinical diagnosis, biochemical analysis, and drug screening. Background technique [0002] Multiple biochemical detection is of great significance for the rapid and efficient diagnosis of biomedical research and clinical diseases. In biomedical research, the development of many diseases involves the synergistic participation of multiple biochemical detection indicators, so it is necessary to simultaneously perform the detection and content analysis of multiple biochemical markers from one sample. For example, cancer marker screening, eight preoperative items (HBV / HCV / HIV / TP, etc.), and respiratory infection pathogens (influenza, pulmonary bronchial, adenovirus), etc. Limited by the extremely high requirements of multiple ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/533G01N33/53G01N33/569G01N33/68
CPCG01N33/5308G01N33/533G01N33/54313G01N33/569G01N33/56983G01N33/68
Inventor 汤曼刘侃陈锦耀艾钊李颂战
Owner WUHAN TEXTILE UNIV
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